不同活化表型的巨噬细胞对Lewis肺癌细胞增殖和侵袭的影响  

Effect of macrophages with different activated phenotype on proliferation and invasion of Lewis lung carcinoma cells

作　　者：章必成[1] 王俊[1] 赵勇[2] 高建飞[2] 郭燕[1] 陈正堂[1] 

机构地区：[1]第三军医大学新桥医院全军肿瘤诊治中心重庆市肿瘤生物技术研究所,重庆400037 [2]广州军区武汉总医院肿瘤科,武汉430070

出　　处：《第三军医大学学报》2007年第11期1013-1016,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30371586)~~

摘　　要：目的探讨不同活化表型的小鼠巨噬细胞对小鼠Lewis肺癌(LLC)细胞增殖和侵袭的影响。方法分别以mr IFN-γ+LPS、mr IL-4处理小鼠RAW264.7巨噬细胞,24h后以RT-PCR鉴定它们表达经典活化的巨噬细胞(caMphi)或替代性活化的巨噬细胞(aaMphi)的标识iNOS或Fizz1的情况。采用Transwell系统共培养活化的巨噬细胞(分为4组:空白对照组、caMphi组、aaMphi组、RAW264.7组)和LLC细胞,绘制LLC细胞生长曲线,并以免疫细胞化学法检测LLC细胞表达血管内皮生长因子-C(VEGF-C)情况,以硝酸还原酶法检测培养液上清中NO含量。分别采用3H-TdR掺入法和Tran-swell侵袭模型观察活化的巨噬细胞对LLC细胞增殖和侵袭的影响。结果成功地建立了小鼠caMphi和aaMphi模型。在4个共培养组中,aaMphi组的LLC细胞生长最快,表达VEGF-C最强,RAW264.7组次之;caMphi组大量分泌NO,其余3组未见NO分泌。通过3H-TdR掺入法和Transwell侵袭模型观察到,aaMphi组的LLC细胞增殖速度最快,侵袭能力最强;RAW264.7组次之;caMphi组的LLC细胞的增殖和侵袭则受到抑制。结论aaMphi能够促进LLC细胞的增殖和侵袭,与其上调后者表达VEGF-C有关;caMphi能够抑制LLC细胞的增殖和侵袭,与其大量分泌NO有关。在与LLC细胞共培养后,RAW264.7细胞的功能朝着aaMphi的方向极化。Objective To explore the effect of mouse macrophages with different activated phenotype on the proliferation and invasion of mouse Lewis lung carcinoma （LLC） cells. Methods RAW264.7 macrophages were treated by mr IFN-γ ＋ LPS or mr IL-4 for 24 h respectively, then identified by RT-PCR to detect iNOS-the marker of classically activated macrophages （caMphi） and Fizzl-the marker of alternatively activated macrophages （aaMphi）. Based on the co-culture Transwell systems of activated macrophages （4 groups： blank control group, caMphi group, aaMphi group and RAW264.7 group） and LLC cells, cell growth curves of LLC cells were drawn, vascular endothelial growth factor-C （VEGF-C） expressions of LLC cells were detected by immunocytochemistry, and NO concentrations of co-culture fluid supernatant were determined by nitrate reductase method. ^3H-TdR incorporation and Transwell invasion model were used to observe the effect of activated macrophages on the proliferation and invasion of LLC cells, respectively. Results Mouse caMphi and aaMphi models were established successfully. Among the 4 co-culture groups, the growth of LLC cells and VEGF-C expression were the fastest and strongest in aaMphi group, secondary in RAW264.7 group; NO concentration was high in caMphi group, but no NO was determined in the other 3 groups. The proliferation and invasion of LLC cells were promoted most in aaMphi group, then in RAW264. 7 group, but inhibited in caMphi group.Conclusion aaMphi can promote the proliferation and invasion of LLC cells by the promotion of VEGF-C expression, caMphi can inhibit them by high level production of NO. After co-culture with LLC cells, the function of RAW264.7 cells may be polarized toward that of aaMphi.

关 键 词：巨噬细胞 活化表型 LEWIS肺癌 增殖 侵袭 

分 类 号：R392.12[医药卫生—免疫学] R730.2[医药卫生—基础医学]